LOCUS       pEF.DEST51-PTPN1        8382 bp DNA     circular SYN 15-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8382)
  AUTHORS   James
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2024 from SnapGene Viewer 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8382
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        475..1653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          706..1644
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1670..1688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1720..1744
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1807..3577
                     /codon_start=1
                     /label=PTPN11
                     /note="Homo sapiens protein tyrosine phosphatase 
                     non-receptor type 11 (PTPN11), transcript variant 1, mRNA 
                     NCBI Reference Sequence: NM_002834.5"
                     /translation="MTSRRWFHPNITGVEAENLLLTRGVDGSFLARPSKSNPGDFTLSV
                     RRNGAVTHIKIQNTGDYYDLYGGEKFATLAELVQYYMEHHGQLKEKNGDVIELKYPLNC
                     ADPTSERWFHGHLSGKEAEKLLTEKGKHGSFLVRESQSHPGDFVLSVRTGDDKGESNDG
                     KSKVTHVMIRCQELKYDVGGGERFDSLTDLVEHYKKNPMVETLGTVLQLKQPLNTTRIN
                     AAEIESRVRELSKLAETTDKVKQGFWEEFETLQQQECKLLYSRKEGQRQENKNKNRYKN
                     ILPFDHTRVVLHDGDPNEPVSDYINANIIMPEFETKCNNSKPKKSYIATQGCLQNTVND
                     FWRMVFQENSRVIVMTTKEVERGKSKCVKYWPDEYALKEYGVMRVRNVKESAAHDYTLR
                     ELKLSKVGQGNTERTVWQYHFRTWPDHGVPSDPGGVLDFLEEVHHKQESIMDAGPVVVH
                     CSAGIGRTGTFIVIDILIDIIREKGVDCDIDVPKTIQMVRSQRSGMVQTEAQYRFIYMA
                     VQHYIETLQRRIEEEQKSKRKGHEYTNIKYSLADQTSGDQSPLPPCTPTPPCAEMREDS
                     ARVYENVGLMQQQK"
     CDS             join(3586..3588,3589..3591,3592..4302)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     protein_bind    complement(4318..4342)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4368..4409
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4419..4436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4465..4689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4735..5163
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5177..5507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5358..5493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5555..5602
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5621..6019
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6177..6298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6749..7337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7508..8299,8300..8368))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(8369..8382,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgcctc gtgaggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1621 caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact aatacgactc
     1681 actataggga gacccaagct ggctaggtaa gcttgatcaa caagtttgta caaaaaagca
     1741 ggctggcgcc ggaaccaatt cagtcgactg gatccggtac cgaggagatc tgccgccgcg
     1801 atcgccatga catcgcggag atggtttcac ccaaatatca ctggtgtgga ggcagaaaac
     1861 ctactgttga caagaggagt tgatggcagt tttttggcaa ggcctagtaa aagtaaccct
     1921 ggagacttca cactttccgt tagaagaaat ggagctgtca cccacatcaa gattcagaac
     1981 actggtgatt actatgacct gtatggaggg gagaaatttg ccactttggc tgagttggtc
     2041 cagtattaca tggaacatca cgggcaatta aaagagaaga atggagatgt cattgagctt
     2101 aaatatcctc tgaactgtgc agatcctacc tctgaaaggt ggtttcatgg acatctctct
     2161 gggaaagaag cagagaaatt attaactgaa aaaggaaaac atggtagttt tcttgtacga
     2221 gagagccaga gccaccctgg agattttgtt ctttctgtgc gcactggtga tgacaaaggg
     2281 gagagcaatg acggcaagtc taaagtgacc catgttatga ttcgctgtca ggaactgaaa
     2341 tacgacgttg gtggaggaga acggtttgat tctttgacag atcttgtgga acattataag
     2401 aagaatccta tggtggaaac attgggtaca gtactacaac tcaagcagcc ccttaacacg
     2461 actcgtataa atgctgctga aatagaaagc agagttcgag aactaagcaa attagctgag
     2521 accacagata aagtcaaaca aggcttttgg gaagaatttg agacactaca acaacaggag
     2581 tgcaaacttc tctacagccg aaaagagggt caaaggcaag aaaacaaaaa caaaaataga
     2641 tataaaaaca tcctgccctt tgatcatacc agggttgtcc tacacgatgg tgatcccaat
     2701 gagcctgttt cagattacat caatgcaaat atcatcatgc ctgaatttga aaccaagtgc
     2761 aacaattcaa agcccaaaaa gagttacatt gccacacaag gctgcctgca aaacacggtg
     2821 aatgactttt ggcggatggt gttccaagaa aactcccgag tgattgtcat gacaacgaaa
     2881 gaagtggaga gaggaaagag taaatgtgtc aaatactggc ctgatgagta tgctctaaaa
     2941 gaatatggcg tcatgcgtgt taggaacgtc aaagaaagcg ccgctcatga ctatacgcta
     3001 agagaactta aactttcaaa ggttggacaa gggaatacgg agagaacggt ctggcaatac
     3061 cactttcgga cctggccgga ccacggcgtg cccagcgacc ctgggggcgt gctggacttc
     3121 ctggaggagg tgcaccataa gcaggagagc atcatggatg cagggccggt cgtggtgcac
     3181 tgcagtgctg gaattggccg gacagggacg ttcattgtga ttgatattct tattgacatc
     3241 atcagagaga aaggtgttga ctgcgatatt gacgttccca aaaccatcca gatggtgcgg
     3301 tctcagaggt cagggatggt ccagacagaa gcacagtacc gatttatcta tatggcggtc
     3361 cagcattata ttgaaacact acagcgcagg attgaagaag agcagaaaag caagaggaaa
     3421 gggcacgaat atacaaatat taagtattct ctagcggacc agacgagtgg agatcagagc
     3481 cctctcccgc cttgtactcc aacgccaccc tgtgcagaaa tgagagaaga cagtgctaga
     3541 gtctatgaaa acgtgggcct gatgcaacag cagaaaagcg gccgcatggt gagcaagggc
     3601 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3661 cacaagttca gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     3721 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     3781 ggctacggcg tggcctgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     3841 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt
     3901 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3961 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4021 ttcaacagcc acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggctaac
     4081 ttcaagatcc gccacaacgt tgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4141 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag
     4201 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4261 accgccgccg ggattacaca tggcatggac gagctgtaca agtagtaaga tatctagacc
     4321 cagctttctt gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc
     4381 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     4441 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4501 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4561 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4621 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4681 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     4741 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4801 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4861 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4921 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4981 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5041 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5101 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5161 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag
     5221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5281 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5341 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5401 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5461 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5521 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     5581 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca
     5641 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     5701 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     5761 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     5821 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     5881 gagcccctgc ggacggtgtc gacaggtgct tctcgatctg catcctggga tcaaagcgat
     5941 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     6001 ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     6061 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6121 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6181 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6241 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6301 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6361 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6421 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6481 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     6961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8341 gggcgacacg gaaatgttga atactcatac tcttcctttt tc
//
